Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT‐LF assay

作者：Qiaozhen Wei, Qiushi Hu, Fengjuan Shi, Shuang Li, Chongsi Sun, Huicong Zhang, Lei Xue, Qiuxia Feng, Jinying Dong, Yongjun Jiao, Lei Zhou · 发表于：Engineering in Life Sciences · 年份：2020 · DOI：10.1002/elsc.202000031 · 被引用次数：5 · 研究领域：Biosensors and Analytical Detection、Escherichia coli research studies、Advanced biosensing and bioanalysis techniques

Abstract Shiga toxin‐producing Escherichia coli (STEC) infection causes a series of diseases that are highly pathogenic and deadly in humans and animals, seriously endangering public health. Of the pathogenic factors within STEC, the two groups of Shiga toxin (Stx) consisting Stx1 and Stx2 plays a prominent role in the pathogenesis of STEC infection. In this study, we developed single‐target up‐converting phosphor technology‐based lateral flow assay (Stx‐UPT‐LFA) for the rapid detection of Stx1 and Stx2, respectively, and also developed a dual‐target Stx1/2‐UPT‐LFA based on single‐target strips to detect of Stx1 and Stx2 at the meantime within 20 min. We choose the purified Stx1 and Stx2 standard samples, and the optimum monoclonal antibody (namely 8E7‐E6, 2F6‐F8 for Stx1 and S1D8, S2C4 for Stx2) were selected for use in Stx‐UPT‐LFA in double‐antibody‐sandwich mode. The sensitivities of single‐target Stx‐UPT‐LFA for both Stx1 and Stx2 were 1 ng mL −1 with accurate quantitation ranges of 1–1000 ng mL −1 and 1–800 ng mL −1 respectively. No false‐negative result was found in the Stx2‐UPT‐LFA even with a high‐test concentration up to 1000 ng mL −1 . Meanwhile, both targets detection sensitivities for dual‐target Stx1/2‐UPT‐LFA were 5 ng mL −1 , and accurate quantitation ranges were 5–1000 ng mL −1 and 5–800 ng mL −1 for standard Stx1 and Stx2 solutions without cross‐interference between two targets. Both techniques showed good linearities, with a linear fitting coefficient of det...